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Anticariogenicity of calcium phosphate complexes of tryptic casein phosphopeptides in the rat.

Casein phosphopeptides (CPP) stabilize calcium phosphate through the formation of casein-phosphopeptide amorphous calcium-phosphate complexes (CPP-CP). The ability of CPP-CP to reduce caries activity was investigated by use of specific-pathogen-free rats inoculated with Streptococcus sobrinus. The animals consumed a defined cariogenic diet free of dairy products. Solutions (100 microL) of the CPP-CP (0.1, 0.2, 0.5, 1.0% w/v) were applied to the animals' molar teeth twice daily. Other groups of animals received solutions containing 500 ppm F, the non-phosphorylated peptides of a casein tryptic digest (0.5% w/v), or the calcium-phosphate complex of a synthetic octapeptide, Ac-Glu-Ser(P)-Ile-Ser(P)-Ser(P)-Ser(P)-Glu-Glu-NHMe, corresponding to the common sequence in the CPP. The CPP-CP significantly reduced caries activity in a dose-response fashion, with 1.0% CPP-CP producing 55% and 46% reductions in smooth surface and fissure caries activity, respectively, being similar to that of 500 ppm F. The anticariogenic effects of CPP-CP and fluoride were additive, since animals receiving 0.5% CPP-CP plus 500 ppm F had significantly lower caries activity than those animals receiving either CPP-CP or fluoride alone. The tryptic digest of casein with the phosphopeptides selectively removed showed no anticariogenic activity. The synthetic octapeptide-calcium phosphate complex significantly reduced caries activity, confirming that this calcium-phosphate-stabilizing portion of the casein phospho-peptides is associated with anticariogenicity. The CPP-CP did not significantly affect the level of S. sobrinus in fissure plaque.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗

Arrest of root surface caries in situ.

This study tests the hypothesis that daily oral hygiene combined with topical fluoride arrests active root-surface caries lesions without changing the mineral content of the lesions. Therefore, changes in mineral content and distribution were studied in root surfaces during caries lesion development and subsequent arrest of lesion progression in situ. In 18 subjects, lesions were developed during 3 months in sound root-surface specimens inserted into lower partial dentures. After 3 months, ground sections were prepared from each lesion prior to re-insertion of the specimens into the dentures. In addition, one sound root specimen was added per subject. During the following 3 months, half of the subjects cleaned both sound and carious specimens once a day with an 1100-ppm fluoride toothpaste, and the specimens were treated twice with 2% NaF for 2 min in situ. The other half of the subjects continued the experiment without cleaning. During the initial three-month period, all specimens developed subsurface lesions extending 187 to 583 microm into the dentin. Lesion depth increased somewhat in both experimental groups during the following 3 months (P > or = 0.1). There was a non-significant increase in mineral loss in the plaque-covered specimens (P = 0.08). However, the total mineral content of specimens subjected to plaque removal and topical fluoride did not change. This treatment resulted in an increased mineral content in the surface layer (P < 0.01) and formation of a zone of higher mineral content within the body of the lesion. The sound root surfaces which had been cleaned for a three-month period showed mineral uptake in the surface layer, occasionally associated with subsurface demineralization extending 20 to 70 microm into the tissue. The mineral loss of these specimens was significantly smaller than that of plaque-covered surfaces (P < 0.001). It is concluded that daily plaque removal and topical fluoride use influence the distribution of mineral in sound and carious root surfaces and may arrest lesion progression without affecting the total mineral content.

Aged↗

Composition of plaque and saliva following a sucrose challenge and use of an alpha-tricalcium-phosphate-containing chewing gum.

Calcium phosphate concentrations in plaque, plaque fluid, and saliva play an important role in caries prevention. In this study, we used a microanalytical technique to examine the anticaries potential of a 2.5% (mass fraction) alpha-tricalcium-phosphate-fortified experimental gum by measuring the pH, free and total calcium, and total phosphate in plaque fluid, whole plaque, and saliva, and centrifuged saliva from 14 subjects who (1) accumulated plaque for 48 hours, (2) fasted overnight, (3) rinsed for 1 min with sucrose, and (4) chewed a control or experimental gum for 15 min. From these data, the hydroxyapatite (HAp) ion activity products (IAP[HAp]) of saliva and plaque fluid were calculated as a measure of tooth mineral saturation. Results, compared with those of the control gum, show significant increases in pH and in free calcium and phosphate concentrations in plaque fluid and saliva when the experimental gum was chewed following sucrose ingestion. These increases result in a rise in fluid saturation with respect to tooth mineral that, for plaque fluid, nearly cancels the decrease seen with the control gum after the sucrose rinse. This suggests that the experimental gum may be more effective than a conventional gum in ameliorating the cariogenic effects of sucrose. Similar statistically significant increases were also seen in the total calcium content of the plaque fluid, centrifuged saliva, whole saliva, and whole plaque, and in the total phosphate of whole plaque and whole saliva. These results suggest that the deposition of a mineral reservoir in plaque and saliva by the experimental gum may help resist future cariogenic challenges.

Adult↗

CO2 laser inhibitor of artificial caries-like lesion progression in dental enamel.

Several studies during the last 30 years have demonstrated the potential of laser pre-treatment of enamel or tooth roots to inhibit subsequent acid-induced dissolution or artificial caries-like challenge in the laboratory. The overall objective of ongoing studies in our laboratories is to determine, systematically, the optimum sets of parameters for carbon dioxide laser irradiation that will potentially effectively inhibit dental caries in enamel and tooth roots. The aim of the present study was to examine the roles of wavelength and fluence in the prevention of caries progression in vitro in enamel by means of a pH-cycling model. The hypothesis to be tested was that the highly absorbed 9.3- and 9.6-microm wavelengths would be efficiently converted to heat, creating a temperature sufficiently high to reduce the acid-reactivity of the mineral and inhibit caries-like lesion progression in dental enamel. One hundred and sixty caries-free tooth crowns were cleaned and varnished with acid-resistant varnish, leaving one exposed window of enamel. Twelve groups of 10 enamel samples were irradiated in their individual windows by one of the four wavelengths (9.3, 9.6, 10.3, or 10.6 microm) of a tunable CO2 laser. Energy per pulse was 25, 50, 100, 200, or 250 mJ (25 pulses). Repetition rate was 10 Hz, and beam diameter was 1.6 mm. Fluence conditions of 1 to 12.5 J/cm2 per pulse were produced. All teeth, including 40 non-irradiated controls, were subjected to pH-cycling to produce artificial caries-like lesions. Results were assessed by cross-sectional microhardness testing. Inhibition of caries progression of from 40% to 85% was achieved over the range of laser conditions tested. At 9.3 and 9.6 microm, 25 pulses at absorbed fluences of 1 to 3 J/cm2 produced inhibition on the order of 70% with minimal subsurface temperature elevation (< 1 degree C at 2 mm depth), comparable with inhibition produced in this model with daily fluoride dentifrice treatments. Safety and efficacy studies will be required in animals and humans before these promising laboratory results can be applied in clinical practice.

Absorption↗

Effects of fluoride-supplemented sucrose on experimental dental caries and dental plaque pH.

Sucrose, 5% and 10% (w/v), supplemented with between 0 and 5 ppm fluoride (F), was tested for its influence in vitro on plaque-induced experimental in vitro enamel caries and plaque pH. Plaque growth on bovine enamel was initiated from saliva inocula and sustained in a multiple plaque growth system for up to 31 days by means of a basal medium with periodic applications of sucrose or sucrose supplemented with F. Change in enamel mineralization was assessed, before and after plaque growth, by microhardness testing and microradiography; pH was monitored with microelectrodes. It was found that enamel demineralization was inversely related to the F concentration in the range 2 to 5 ppm, for both 5% and 10% sucrose. Plaque pH responses were unaffected by the F supplements.

Analysis of Variance↗

Animal caries models for evaluating fluoride dentifrices.

To determine the ability of animal caries models to evaluate the cariostatic potential of fluoride dentifrices, we examined four different coronal caries models. These currently used rat caries models were: Francis' hypomineralized area model, Gaffar's CARA rat model, the Connecticut model, and the Indiana model. Available data indicated that all of these models were capable of detecting significant cariostatic benefits from clinically proven fluoride dentifrices. For the most part, these animal models were also able to demonstrate a response to increasing concentrations of fluoride, particularly if the fluoride was provided in an ionic rather than a complexed form. Data were also presented which indicated the potential for animal caries models to be used to predict the clinical benefits of non-fluoride systems as well as the utility of fluoride systems for the prevention of root-surface caries. It is concluded that animal caries models play a significant role in the development and evaluation of fluoride and other systems for the clinical prevention of dental caries.

Animals↗

In situ caries models.

By using in situ models, we have the potential to study both fundamental aspects of the caries process as well as more applied research problems such as the effect of food on dental caries and the role of fluoride in caries prevention in human subjects without actually causing caries in the natural dentition. The key experimental parameters that need to be considered in the development of an in situ model are the characteristics of the subject panel, the physical design of the model, the type of hard tissue substrate and the method of assessing mineral status, and the study design and clinical protocol. Each parameter must be carefully considered in relation to the objectives of the research, study design requirements, ethical considerations, impact on clinical relevance, and impact on the control of variation. The major source of variation associated with in situ models should be of biological and not experimental origin. The design and conduct of proper in situ model studies require a clear understanding of the caries process, sound analytical support, and a knowledge of how to work with research subjects to achieve a high level of compliance. Given the complex nature of caries, a combination of hard tissue substrates--including sound, surface-softened lesions and subsurface lesions--may be necessary to model all aspects of caries progression and prevention successfully. Internal validation of in situ models using fluoride dose-response controls is considered to be necessary for studies evaluating the efficacy of new fluoride dentifrice formulations.

Animals↗

The application of in situ models for evaluation of new fluoride-containing systems.

Many in situ models have assessed the anticaries potential of fluoride-containing systems (Stookey et al., 1985; Mellberg et al., 1986, 1992a,b; Corpron et al., 1986; Featherstone and Zero, 1992; Ogaard and Rolla, 1992; Stephen et al., 1992). Several models have reportedly been validated according to guidelines proposed by Proskin et all. (1992). The proposed guidelines cover only dentifrices containing sodium fluoride (NaF) or sodium monofluorophosphate (SMFP) as active ingredients. These compounds are the most widely used sources of fluoride in dentifrices, and dose-response clinical standards are available for both. Other fluoride compounds, such as amine fluoride (AmF) and stannous fluoride (SnF2), have also been proven effective in reducing caries (Muehler et al., 1957, 1958; Marthaler, 1968; Lu et al., 1980; Cahen et al., 1982). Profile standards for these fluorides were not included in the proposed guidelines, primarily due to the lack of clinical data necessary to establish a dose response for these ingredients. Criteria for demonstrating the efficacy of these ingredients, along with methods to assess new fluoride compounds, need to be established. In situ models are used to evaluate the anticaries potential of new compounds added to mouthrinses, gums, slow-release devices, etc. (Creanor et al., 1992; Manning and Edgar, 1992; Lamb et al., 1993; Toumba and Curzon, 1993; Wang et al., 1993). Ingredients are often added to dentifrices previously proven effective against caries in order to provide additional benefits of gum health, tartar control, cleaning, etc. Proposals are made regarding the in situ testing of new dentifrices containing clinically proven fluoride compounds other than NaF and SMFP, as well as alternative delivery systems, in order to assist in their evaluation.

Ampholyte Mixtures↗

Reproducibility of TMR for the determination of longitudinal mineral changes in dental hard tissues.

Transverse microradiography (TMR) of thin sections is the principal method for determining mineral changes in experimental lesions. Because it has become also the standard by which newly-developed methods are validated, demands on all steps of the procedure are high. Thin sections of enamel and dentin require high-precision cutting or lapping to preserve surface structures, but their different brittleness and elasticity preclude a uniform procedure. Image analysis permits the measurement of fine details at a micron scale, but because most studies are aimed at overall mineral changes, they are usually lost on broadening of the scan areas and averaging of experimental groups. Small errors in repositioning of the scan areas for repetitive measurements may yield distorted lesion profiles, while quantitative data such as the integrated mineral loss are hardly influenced. A major reason for "irreproducibility" of TMR are the different definitions of the lesion parameters as used by various research groups. We recommend that these definitions be agreed on if the full potential of TMR as an international standard is to be utilized.

Absorptiometry, Photon↗

An abbreviated caries clinical trial design validated over 24 months.

Conventional caries trials last from 24 to 36 months. This study evaluated whether the previously established difference in efficacy between 1000- and 2500-ppm-fluoride dentifrices could be detected after 12 months. Caries was assessed by clinical visual assessment (CVA-simplified version of Dundee Selectable Threshold Method - DSTM), bitewing radiography, and Fiber Optic Transillumination (FOTI). Changes in status for individual surfaces were classified by means of pre-prepared matrices as 0 (unchanged), +1 (initiation or progression), or -1 (regression) and summed for each subject to yield an event score. Mean group event scores were calculated for each product. DSTM at the D(1) [enamel and dentin] threshold showed significant inter-group differences in mean event scores (p < 0.003) and D(1)MFS increment (< 0.007) at 12 months; these were confirmed at 24 months by traditional increment analysis (CVA & FOTI at the D(3) (dentin only) threshold + radiography, p < 0.03). This study confirms the validity of an abbreviated trial protocol.

Adolescent↗

Improving clinical visual detection--potential for caries clinical trials.

It has been the norm in caries clinical trials to measure caries increment by several different caries-recording systems, including the crude DMF-S/T index. However, there is a reserved attitude as to whether to subdivide the non-cavitated lesions and use arrested lesions in clinical trials. This has been due to the belief that it is not possible to achieve reliable data of the early stages of the disease (Radike, 1972). However, recently, Ekstrand et al.(1997, 1998) showed that it was possible: (1) to differentiate between different stages of non-cavitated occlusal lesions, (2) to differentiate between active and inactive occlusal lesions, and (3) to predict the depth of the lesion. In at least 4 other clinical studies, the reproducibility of recording initial active lesions, cavitated active lesions, and arrested lesions was found to be adequate (Carvalho et al., 1989; Nyvad et al, 1999; Ekstrand et al., 2000; Machiulskiene et al., 2001). Since caries today is a more slowly developing disease in many countries in the world, this will result in prolongation of the duration of the clinical trial, which will increase the costs. As indicated above, there is now sufficient evidence that caries can be clinically diagnosed accurately and reliably in earlier stages as well as in an arrested stage. If such stages of caries are used as outcome variables in caries clinical trials, they may have a positive influence on the trials' duration and costs.

Clinical Trials as Topic↗

Tensile strength of mineralized/demineralized human normal and carious dentin.

The bond strengths of resins to caries-affected dentin are low. This could be due to weakened organic matrix. The purpose of this work was to determine if the ultimate tensile strength (UTS) of excavated carious dentin is weaker than that of normal dentin. Soft caries was excavated from extracted human molars, and the tooth was vertically sectioned into slabs. Each slab was trimmed to an hourglass shape, parallel or perpendicular to the tubule direction. Half of the specimens were mineralized, while the other half were completely demineralized in EDTA. ANOVA on ranks showed that the three-factor interactions (mineralization, caries, tubule direction) were all significant (p < 0.0001), indicating that mineralization and tubule direction gave different UTS results in normal and caries-affected dentin. No significant differences were seen between the UTS of normal and and that of caries-affected demineralized dentin in the parallel or perpendicular group. The matrix of demineralized caries-affected dentin was as strong as that of normal demineralized dentin when tested in the same direction.

Chelating Agents↗

Ca, Pi, and F in the fluid of biofilm formed under sucrose.

Calcium (Ca), inorganic phosphorus (P(i)), and fluoride (F) concentrations are low in the whole plaque biofilm formed under exposure to sucrose. It was hypothesized that this would be reflected in the biofilm fluid, where these low values should greatly influence the de/remineralization process. Dental biofilms were formed in situ over enamel blocks mounted in palatal appliances and exposed 8 times/day to distilled water, glucose+fructose, or sucrose solutions for 14 days. While Ca, P(i), and F concentrations in the whole biofilms were significantly lower in the glucose+fructose and sucrose groups, no effect on biofilm fluid was observed, even after a cariogenic challenge. An increase in whole biofilm mineral ions was observed 24 hrs after the carbohydrate treatments were suspended, but this effect was also not observed in the fluid. These results suggest that there is a homeostatic mechanism that maintains biofilm fluid mineral ion concentration, regardless of its total concentration in the whole biofilm.

Adult↗

An investigation into the effect of a fluoride releasing adhesive on the prevalence of enamel surface changes associated with directly bonded orthodontic attachments.

Despite careful patient selection, decalcification still remains a problem during fixed orthodontic treatment. The advantages of a method of delivering fluoride to the area of the tooth most at risk, which does not rely on patient compliance, makes a fluoride releasing bonding adhesive desirable. The aims of this study were to investigate the potential of a fluoride releasing bonding material for preventing decalcification and to monitor its efficacy at retaining orthodontic brackets. In order to carry out the first part of the study it was necessary to devise a reproducible and sensitive method of recording the extent and severity of any decalcification. A photographic technique which fulfils these criteria is described.

Adhesives↗

Caries prevention: current strategies, new directions.

Despite major advances in caries prevention, a large subset of the U.S. population still needs additional control measures. In numerous other countries, needs are escalating as caries rates continue to rise. Building on current strategies and new insights into the specific mechanisms of caries initiation, researchers are creatively using a variety of new technologies, especially in molecular biology, to fashion a new generation of preventive measures.

Anti-Infective Agents, Local↗

Caries prevention in the 21st century.

While the incidence of caries has decreased during the past 50 years because of the introduction of water fluoridation and fluoride toothpastes, it is still widespread. Improved therapies, using topical treatments to replace lost calcium and phosphate minerals from early carious lesions, will reduce the need for surgical intervention in the future. Early caries detection techniques currently being developed will allow dentists to maximize the usefulness of these treatments.

Adolescent↗

The effects of seven carbamide peroxide bleaching agents on enamel microhardness over time.

BACKGROUND: Different concentrations of carbamide peroxide (10 to 22 percent) have been used successfully as bleaching agents, but the adverse effects on enamel microhardness at different times are unknown. METHODS: The authors analyzed seven bleaching agents and a placebo. The agents were applied on the surface of human dental fragments for eight hours per day for 42 days and stored in vials containing artificial saliva. Microhardness measurements were performed at baseline, eight hours and seven, 14, 21, 28, 35 and 42 days, as well as at seven and 14 days after treatment had ended. RESULTS: Enamel treated with different bleaching agents or a placebo experienced a similar decrease in microhardness values over time, with the exception of fragments exposed to Opalescence PF 20 percent (Ultradent Products, South Jordan, Utah), or OPA20. Until the 49th day, the enamel exposed to OPA20 exhibited the lowest differences from baseline values. After 14 posttreatment days, enamel treated with placebo, Nite White 10 percent Excel (Discus Dental, Los Angeles), Nite White 16 percent Excel and Opalescence 10 percent exhibited the greatest differences from baseline values. An increase in enamel microhardness occurred at the end of the posttreatment period, although baseline values were not reached. CONCLUSIONS: Different concentrations of carbamide peroxide agents result in decreases in enamel microhardness. A post-bleaching period in artificial saliva resulted in recovery of baseline microhardness values (one agent) or an increase in values, although baseline values were not reached for these products. Clinical Implications. Higher concentrations of carbamide peroxide containing 0.11 percent ion fluoride can bleach teeth in a shorter period, with fewer hazardous effects on the enamel mineral content.

Carbamide Peroxide↗